Premium
A high‐throughput microchip‐based glycan screening assay for antibody cell culture samples
Author(s) -
Primack Jeremy,
Flynn Gregory C.,
Pan Hai
Publication year - 2011
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201000619
Subject(s) - glycan , chromatography , monoclonal antibody , cell culture , clone (java method) , antibody , microbiology and biotechnology , chemistry , cell , high throughput screening , biology , biochemistry , glycoprotein , immunology , dna , genetics
A high‐throughput screening assay was developed to quantify major glycan species in the crude mammalian cell culture samples for monoclonal antibodies (mAbs). This method utilizes high‐speed microchip electrophoresis separation following a fast sample preparation procedure. Using a 96‐well ultra‐filtration membrane, interfering species in the cell culture media were efficiently removed as the samples were concentrated. A commercial microchip electrophoresis instrument was used for high‐speed separation, allowing each sample to be analyzed in less than 1 min. This method is well suited for the purpose of high‐throughput antibody glycan profiling during cell culture expression, including clone selection and cell culture process optimization. The relative levels of high mannose (HM), fucosylated and galactosylated glycan species in the Fc domain can be determined for hundreds of crude cell culture samples in a few hours.